Monitoring protein kinase activity in cell lysates using a high-density peptide microarray

Xiaoming Han, Go Yamanouchi, Takeshi Mori, Jeong Hun Kang, Takuro Niidome, Yoshiki Katayama

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Monitoring and targeting protein kinases is widely accepted as a promising approach for disease diagnosis and drug discovery. For this purpose, the authors have developed an original type of peptide array as a high-throughput screening assay for quantitatively evaluating kinase activity. A volume of 2 nL of peptide solution was spotted onto a formyl group-modified glass slide by using an arrayer, which was designed for use with protein chip technology. The phosphorylation was recognized by fluorescence-label antibody and detected with an automatic microarray scanner widely used in DNA chip technology. The system needs low sample volume, provides a high-density peptide array, and supplies high reproducibility. It provided enough sensitivity for inhibitor screening, even though a relatively low concentration of purified kinase was employed. The assay also proved useful for the detection of intracellular kinase activity as well as for the measurement of the fluctuations of intracellular protein kinase activity with drug stimulation. Thus, this peptide array would be applicable for kinase-targeted diagnosis, cell-based drug screening, and signal pathway investigation.

Original languageEnglish
Pages (from-to)256-262
Number of pages7
JournalJournal of Biomolecular Screening
Volume14
Issue number3
DOIs
Publication statusPublished - Mar 1 2009

Fingerprint

Microarrays
Protein Kinases
Phosphotransferases
Screening
Peptides
Monitoring
Assays
High-Throughput Screening Assays
Technology
Protein Array Analysis
Preclinical Drug Evaluations
Phosphorylation
Drug Discovery
Oligonucleotide Array Sequence Analysis
Pharmaceutical Preparations
Glass
Labels
Signal Transduction
Fluorescence
Throughput

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

Cite this

Monitoring protein kinase activity in cell lysates using a high-density peptide microarray. / Han, Xiaoming; Yamanouchi, Go; Mori, Takeshi; Kang, Jeong Hun; Niidome, Takuro; Katayama, Yoshiki.

In: Journal of Biomolecular Screening, Vol. 14, No. 3, 01.03.2009, p. 256-262.

Research output: Contribution to journalArticle

Han, Xiaoming ; Yamanouchi, Go ; Mori, Takeshi ; Kang, Jeong Hun ; Niidome, Takuro ; Katayama, Yoshiki. / Monitoring protein kinase activity in cell lysates using a high-density peptide microarray. In: Journal of Biomolecular Screening. 2009 ; Vol. 14, No. 3. pp. 256-262.
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